| 摘要: |
| ［摘要］ 目的 制备甲状腺素运载蛋白（transthyretin，TTR）的时间分辨免疫荧光（time-resolved fluoroimmunoassay，TRFIA）试剂盒并对其性能进行评价。方法 将抗TTR的4H2单克隆抗体作为包被抗体包被至96孔板，用Eu3+标记3E4单克隆抗体作为检测抗体，制备双抗体夹心TRFIA试剂盒，应用该试剂盒对42例川崎病（Kawasaki disease,KD）患儿及13名健康儿童的血清进行检测，以稀释回收率、准确度、精密度、稳定性等指标对试剂盒的效能进行评价。结果 该研究制备的TTR TRFIA试剂盒与Western blot法同时检测42份KD临床样本和13份健康样本，两方法的结果符合率为100%，特异性强。试剂盒的最低检出浓度为0.05 μg/ml，稀释回收率为88.00%～111.00%，分析内精密度为8.01%～9.17%，分析间精密度为8.88%～11.82%。稳定性实验表明该试剂盒可在4 ℃稳定保存6个月，37 ℃稳定保存7 d。结论 该研究制备的TTR TRFIA试剂盒具有准确性高、方便快捷等优点，适用于大批量临床KD血清样品的TTR检测，为快速区分丙种球蛋白敏感型和无反应型的KD患儿提供了方法。 |
| 关键词: 甲状腺素运载蛋白 川崎病 时间分辨免疫荧光分析 |
| DOI：10.3969/j.issn.1674-3806.2020.01.08 |
| 分类号:R 446 |
| 基金项目:广东省科技计划项目（编号：2016A010119067） |
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| Preparation and effectiveness evaluation of time-resolved fluoroimmunoassa kit for detection of transthyretin |
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CHEN Cui-cui, LIANG Huan-kun, LI Jie-xing, et al.
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Guangzhou Youdi Bio-Technology Co., Ltd., Guangdong 510663, China
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| Abstract: |
| ［Abstract］ Objective To prepare a kit of time-resolved fluoroimmunoassa(TRFIA) for detection of transthyretin(TTR), and evaluate its effectiveness. Methods The clone 4H2, the anti-TTR monoclonal antibody, was coated into 96-well microplate as the capture antibody, and the monoclonal antibody(clone 3E4) labeled with the Eu3+ as the detection antibody. A double-antibody sandwich TRFIA kit was prepared by sandwiching antigens to be tested between two detection antibodies. Then, the kit was used to detect the TTR levels of 42 children with Kawasaki disease(KD) and 13 healthy children, and the efficiency of the kit was evaluated using its indexes of dilution recovery, accuracy, precision and stability. Results The TTR TRFIA kit and Western blot method were adopted to detect the same 42 KD samples and 13 healthy samples at the same time, and the coincidence rate between the two methods was 100%, with a high specificity. The minimum detectable concentration of the prepared TRFIA kit was 0.05 μg/ml with the dilution recovery rate being 88.00%～111.00%, and the intra-assay precision being 8.01%～9.17%, and the inter-assay precision being 8.88%～11.82%. The stability test showed that the kit could be stably stored at 4 ℃ for 6 months and 37 ℃ for 7 days. Conclusion The TTR TRFIA kit prepared in this study has the advantages of high accuracy, convenience and quickness, which is suitable for TTR detection of a large amount of clinical KD serum samples, and provides a new method for differentiation of intravenous immunoglobulin G(IVIG)-sensitive and IVIG-non-responsive KD children. |
| Key words: Transthyretin(TTR) Kawasaki disease(KD) Time-resolved fluoroimmunoassa(TRFIA) |
